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ChemiScreen™ Membrane Prep Human Recombinant MT2

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货号：HTS106M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 MT2
Description
Catalogue Number HTS106M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ Membrane Preparation Recombinant Human MT2 Melatonin Receptor
Overview Full-length human MTNR1B cDNA encoding MT2
Background Information Melatonin (5-methoxy-acetyltryptamine) is a hormone synthesized and secreted in a circadian manner with high levels occurring in all species at night. Melatonin has been implicated in the regulation of various neural and endocrine processes that are cued by the daily change in photoperiod (Masana and Dubocovich, 2001) Two mammalian melatonin receptor subtypes, MT1 and MT2 have been cloned. Both receptors are members of the superfamily of putative seven transmembrane G-protein coupled receptors, which are coupled to the inhibition of cyclic AMP synthesis upon agonist binding. Pharmacological studies suggest that the phase shifting effect of melatonin on circadian rhythms may be mediated by the MT2 receptor subtype (Dubocovich et al. 1998). Melatonin inhibition of dopamine release from the retina is also mediated by the MT2 receptor subtype (Dubocovich et al. 1997). Chemicon's MT2 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of agonists and antagonists at MT2. The membrane preparations exhibit a Kd of 0.1 nM for [125I]-Iodomelatonin. With 0.2 nM [125I]-Iodomelatonin, 5 µg/well MT2 Membrane Prep typically yields greater than 6-fold signal-to-background ratio.
References
Product Information
Format Membranes
Presentation Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to 1 mg/mL in packaging buffer, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80°C.
Applications
Application Human MT2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay and GTPγS binding
Biological Information
Entrez Gene Number
Entrez Gene Summary This gene encodes one of two high affinity forms of a receptor for melatonin, the primary hormone secreted by the pineal gland. This gene product is an integral membrane protein that is a G-protein coupled, 7-transmembrane receptor. It is found primarily in the retina and brain although this detection requires RT-PCR. It is thought to participate in light-dependent functions in the retina and may be involved in the neurobiological effects of melatonin.
Gene Symbol
  • MTNR1B
  • Mel-1B-R
  • MT2
  • MEL-1B-R
GPCR Class A
Host Cells Chem-1
Protein Target MT2
Target Sub-Family Melatonin
UniProt Number
UniProt Summary FUNCTION: SwissProt: P49286 # High affinity receptor for melatonin. Likely to mediates the reproductive and circadian actions of melatonin. The activity of this receptor is mediated by pertussis toxin sensitive G proteins that inhibit adenylate cyclase activity.
SIZE: 362 amino acids; 40188 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed in retina and less in brain and hippocampus.
SIMILARITY: SwissProt: P49286 ## Belongs to the G-protein coupled receptor 1 family.
Incubation Conditions Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 500mM NaCl. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, filtered and stored at 4°C
Radioligand: [125I] Iodomelatonin (Perkin Elmer#: NEX-236)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 6-fold signal:background with [125I]-labeled Iodomelatonin at 0.2 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
2 Table 1. Signal:background and specific binding values obtained in a competition binding assay with MT membrane prep and unlabeled melatonin.
Product Usage Statements
5 µg/well
Signal:Background 9.96
Specific Binding (cpm) 1424.1

SPECIFICATIONS: 1 unit = 5 µg
Bmax for [ 125I]-Iodomelatonin binding: 0.04 pmol/mg protein
Kd for [ 125I]-Iodomelatonin binding: ~0.1 nM Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size 200 units
Transport Information
Supplemental Information
Specifications

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